KR102088113B1 - Skin whitening composition comprising brassinin as active ingredient - Google Patents

Abstract

The present invention relates to a composition for skin whitening comprising brassinin. The composition comprising brassinin according to the present invention suppresses production of melanin and inhibits the activity of tyrosinase, a regulatory enzyme for melanin production, thereby having an excellent effect of skin whitening. Therefore, the composition can be usefully used in cosmetics and food fields.

Description

Skin whitening composition comprising brassinin as active ingredient

The present invention relates to a composition for skin whitening comprising bracinin.

Melanin present in vivo acts as a major biological pigment that changes the color of the skin. Melanosomes and melanin are produced by melanocytes in the basal layer of the skin, which are produced by cells called melanosomes. Various mechanisms are involved in melanin biosynthesis of the skin, including the regulation of alpha-melanosite-regulating hormones produced by the p53 gene by UV-stimulation and the increased microphthalmia associated transfer factor (MITF). Catalyzes the activation of tyrosinase present in the melanosome, initiates the hydroxylation process of DOPA by activated tyrosinase, and promotes melanin biosynthesis by tyrosinase sub-proteins TRP1 and TRP2. And control. The final product, melanin, enters the epidermal cells and serves to protect skin cells from skin irritants such as ultraviolet light. However, over-stimulation of melanin synthesis causes mechanisms such as hyperpigmentation and inflammatory deposits on the skin.

If the melamine-forming melanosites in human skin are not uniformly distributed for any reason, pigment spots are created that are brighter or darker than the surrounding areas of the skin. In order to solve this problem, a whitening agent is used which at least partially makes such pigment spots uniform. Many skin lightening agents contain rather potent tyrosinase inhibitors, which are mainly hydroquinones, for example hydroquinone derivatives such as arbutin, vitamin C, for example ascorbic acid such as ascorbyl palmitate. Derivatives, kojic acid, for example kojic acid derivatives, such as kojic acid dipalmitate, are particularly used as commercially useful skin and hair whitening agents.

Among them, vitamin C and ascorbic acid derivatives do not directly act as tyrosinase inhibitors, so the whitening effect is insufficient, and kojic acid may cause skin allergy.

Accordingly, the present inventors have confirmed that brassine (Brassinin) has an excellent effect on skin whitening by inhibiting melanin production and inhibiting the activity of tyrosinase, a regulation enzyme of melanin production, and completed the present invention.

It is an object of the present invention to provide a cosmetic composition for skin whitening comprising brassinin or an acceptable salt thereof.

Another object of the present invention to provide a health food composition for skin whitening comprising brassin (Brassinin) or an acceptable salt thereof.

Still another object of the present invention is to provide a nutraceutical composition for skin whitening comprising brassinin or an acceptable salt thereof.

Another object of the present invention is to provide a composition for skin whitening comprising Brassinin or an acceptable salt thereof.

In order to achieve the above object,

The present invention provides a cosmetic composition for skin whitening comprising brassine (Brassinin) represented by the following general formula (1) or an acceptable salt thereof:

[Formula 1]

.

.

In another aspect, the present invention provides a health food composition for skin whitening comprising brassine (Brassinin) represented by the formula (1) or an acceptable salt thereof.

Furthermore, the present invention provides a health functional food composition for skin whitening comprising brassine (Brassinin) represented by Formula 1 or an acceptable salt thereof.

Furthermore, the present invention provides a composition for skin whitening comprising brassine (Brassinin) represented by Formula 1 or an acceptable salt thereof.

The composition comprising brassinin according to the present invention has an excellent effect on skin whitening by inhibiting the production of melanin and inhibiting the activity of tyrosinase, a regulatory enzyme of melanin production, and thus may be usefully used in cosmetics and food fields.

Figure 1a shows the chemical structure of the bracinin.
Figure 1b is a result of analyzing the cytotoxicity by treating the bracinin to B16F10 cells.
Figure 1c is a result of analyzing the cytotoxicity by treating the bracinin to HaCaT cells.
Figure 2 shows the tyrosinase inhibitory effect of brassinin.
Figure 3a shows the effect of inhibiting melanin production by bracinin (Intracellular melanin contents: the amount of melanin present in melanocytes).
Figure 3b shows the inhibitory effect of melanin production by bracinin (Extracellular melanin contents: the amount of melanin secreted out of melanocytes).

Hereinafter, the present invention will be described in detail.

Cosmetic composition for skin whitening

The present invention provides a cosmetic composition for skin whitening comprising brassine (Brassinin) represented by the following Chemical Formula 1 or an acceptable salt thereof.

[Formula 1]

.

.

In one embodiment of the present invention, brassinin (Brassinin) or an acceptable salt thereof is characterized in that it has the inhibitory activity of melanin production and tyrosinase.

As used herein, the term "whitening" refers to inhibiting, inhibiting or alleviating hyperpigmentation of the skin. Hyperpigmentation of the skin includes freckles, blemishes, hyperpigmentation after UV exposure, hyperpigmentation after inflammation, aging black spots, brown spots or blotch.

In the cosmetic composition according to the present invention, the cosmetic composition for skin whitening may contain brassinin (Brassinin) of Formula 1 or an acceptable salt thereof in an amount of 0.0001 to 90% by weight based on the total weight of the composition, Preferably in an amount of 0.0001 to 10% by weight.

As used herein, the term "effective amount" means the amount of a compound that can inhibit, inhibit or alleviate hyperpigmentation of the skin. The effective amount of the compound included in the skin whitening composition of the present invention will vary depending on the form in which the skin whitening composition is commercialized, the method in which the compound is applied to the skin, and the time to stay in the skin. For example, when the composition for skin whitening is commercialized as a medicament for dermatological treatment due to hyperpigmentation of the skin, the composition of brassinin or brassinin at a higher concentration than when it is commercialized to cosmetics that are routinely applied to the skin And acceptable salts thereof. Even in the case of cosmetics, in the case of a wash-off type cosmetic such as a makeup remover, a detergent, etc., in which an active ingredient stays on the skin within a short time, the compound may include a relatively high concentration. On the other hand, in the case of a leave-on type cosmetic such as a lotion, an emulsion, a cream, an essence, etc., in which the active ingredient stays on the skin for a long time, a lower concentration of Brassinin or a wash-off type cosmetic is used. It may also include acceptable salts thereof.

The cosmetic composition for skin whitening according to the present invention is a skin external ointment, cream, supple cosmetics, nourishing cosmetics, pack, essence, hair tonic, shampoo, conditioner, hair conditioner, hair treatment, gel, skin lotion, skin softener, skin toner , Astringent, Lotion, Milk Lotion, Moisture Lotion, Nutrition Lotion, Massage Cream, Nutrition Cream, Moisture Cream, Hand Cream, Foundation, Nutrition Essence, Sunscreen, Soap, Cleansing Foam, Cleansing Lotion, Cleansing Cream, Body Lotion and Body Cleanser It may have a formulation selected from the group consisting of, but is not limited thereto. The composition of each of these formulations may contain a variety of bases and additives necessary for the formulation of the formulation and are suitable, and the type and amount of these components can be readily selected by those skilled in the art.

The composition of the present invention may contain one or more skin lightening active ingredients which exhibit the same or similar function in addition to the brassinin of the present invention or an acceptable salt thereof. Skin whitening active ingredients include kojic acid and derivatives thereof, arbutin, ascorbic acid and derivatives thereof, hydroquinone and derivatives thereof, resorcinol, cycloalkanone, methylenedioxyphenyl alkanol, 2,7-dinitroindazole or Plant extracts such as vine tangerine extract, rice extract, licorice extract, and the like, but are not limited thereto.

When the formulation of the present invention is a paste, cream or gel, animal carriers, plant fibers, waxes, paraffins, starches, tracantes, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicas, talc or zinc oxide, etc. may be used as carrier components. Can be.

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used, in particular in the case of a spray, additionally chlorofluorohydrocarbon, propane Propellant such as butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solvating agent or emulsifying agent is used as the carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 Fatty acid esters of, 3-butylglycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan.

When the formulation of the present invention is a suspension, liquid carrier diluents such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters, microcrystalline Cellulose, aluminum metahydroxy, bentonite, agar or tracant and the like can be used.

When the formulation of the present invention is a surfactant-containing cleansing, the carrier component is aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide. Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, linolin derivatives or ethoxylated glycerol fatty acid esters and the like can be used.

Formulations of the present invention may further contain excipients, including fluorescent substances, fungicides, hydrogenic agents, moisturizers, fragrances, fragrance carriers, proteins, solubilizers, sugar derivatives, sunscreens, vitamins, plant extracts, and the like. .

When the cosmetic composition for skin whitening is formulated as a medicine or cosmetic as described above, it may include a known excipient applicable to the skin to act as a carrier for the active ingredient. When formulated as a drug, reference may be made to the contents disclosed in [Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA], and when formulated as a cosmetic, [International cosmetic ingredient dictionary, 6th ed., The cosmetic, Toiletry] and Fragrance Association, Inc., Washington, 1995]. The documents are included as part of this specification.

Health food or functional food composition for skin whitening

The present invention provides a health food or health functional composition for skin whitening comprising brassine (Brassinin) represented by the following Chemical Formula 1 or an acceptable salt thereof.

[Formula 1]

.

.

When using brassine (Brassinin) or an acceptable salt thereof of the present invention as a nutraceutical and health food composition, there is no particular limitation on the type of food. Examples of foods to which the composition of the present invention may be added include drink, meat, sausage, bread, biscuit, rice cake, chocolate, candy, snack, confectionary, pizza, ramen, other noodles, gum, dairy products including ice cream, various Soups, beverages, alcoholic beverages and vitamin complexes, dairy products and dairy products, and the like, and include both nutraceuticals and health food compositions in the conventional sense.

In one embodiment of the present invention, the total composition content may include 0.0001 to 10% by weight of the bracinin or an acceptable salt thereof, but is not limited thereto.

Nutraceuticals and health food compositions containing Brassinin or an acceptable salt thereof according to the present invention can be added as is to foods or used with other foods or food ingredients, and can be suitably used according to conventional methods. have. The mixing amount of Brassinin or an acceptable salt thereof may be suitably determined depending on the purpose of use (prevention or improvement). In general, the amount of the composition in the nutraceutical and health food can be added to 0.1 to 90 parts by weight of the total food weight. However, in the case of prolonged intake for the purpose of maintaining health or controlling the health, the amount may be below the above range, and since there is no problem in terms of safety, brassinin or an acceptable salt thereof is above the above range. Can also be used in amounts.

The nutraceutical and the health food composition of the present invention is not particularly limited to other ingredients except for containing the present invention brassinin (Brassinin) or an acceptable salt thereof as essential ingredients in the indicated ratios, and various flavoring agents such as ordinary beverages. Or natural carbohydrate or the like as an additional component. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And conventional sugars such as polysaccharides such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The ratio of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g per 100 nutraceutical and health food composition of the present invention.

In addition to the above, the functional foods and health food compositions containing brassinin or an acceptable salt thereof of the present invention may be used in various nutrients, vitamins, minerals (electrolytes), synthetic flavors, and natural flavoring agents, such as colorants. And neutralizers (cheese, chocolate, etc.), pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks, and the like. can do. In addition, the nutraceutical and health food composition of the present invention may contain the flesh for the production of natural fruit juice and fruit juice drinks and vegetable drinks.

These components can be used independently or in combination. The proportion of such additives is not so critical but is generally selected in the range of 0.1 to about 20 parts by weight per 100 parts by weight of the nutraceutical and health food composition containing Brassinin or an acceptable salt thereof.

Skin Whitening Composition

The present invention provides a composition for skin whitening comprising brassine (Brassinin) represented by the following formula (1) or an acceptable salt thereof.

[Formula 1]

.

.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following examples are merely to illustrate the present invention, the contents of the present invention is not limited by the following examples.

1. Reagents and Materials

The cell culture plate used in this experiment was a product of SPL, α-MSH (alpha-melanocyte-stimulating hormone), DMEM (Dulbecco's Modified Eagle's Medium-High Glucose), DPBS (Dulbecco's Phosphate Buffered Saline), Tryrosinase from mushroom, L-DOPA, Arbutin, Niacinamide, Brassinin were used by Sigma Aldrich Korea, and FBS was used by ATCC.

1-1. Cell culture

B16F10 cells were distributed by the Korea Cell Line Bank. DMEM with 10% heat inactivated FBS, 100 units / ml and 1% penicillin / streptomycin was incubated in an incubator at 37 ° C and 5% CO ₂ .

1-2. Cell viability assay

Cell viability was measured using Crystal violet (Sigma aldrich) solution. Dispense 2 ml of each well at 2.5 x 10 ⁵ / ml in a 6 well plate, stabilize the plate for 24 hours, and then treat Brassinin at 10, 20, and 40 μM concentrations in the experimental groups, except for the control group. Incubated for days. The cultured cells were fixed in 4% paraformaldehyde solution (Paraformaldehyde solution) for 10 minutes after washing with PBS, and then stained for 10 minutes using 0.5% crystal violet solution.

1-3. External & Intra-Melanin Content Assay

Cell culture was carried out in the same manner as the cell viability measurement, but a medium without phenol red was used. The amount of extracellular melanin content was measured at OD405 nm by the melanin secreted in the cell culture medium, and the intracellular melanin content was measured using 1N NaOH solution. After dissolving melanin for 1 hour at 80 ℃ was measured by absorbance of OD405 nm.

1-4. In vitro Tyrosinase Activity Assay

In vitro Tyrosinase activity was measured using the method of Yagi et al. (1986). Equal amounts of 0.5 ml of 0.175 M sodium phosphate buffer (pH 6.8) and 0.2 ml of 10 mM L-DOPA were added to all test tubes (6). Next, arbutin (1 mM), niacinamide (50 uM) and brassinin (20, 40 uM) were added as experimental groups. 0.2 ml of mushroom tyrosinase (100 U / mL) isolated from mushrooms was added to five test tubes (black graph) except for the control group (white graph). The solution to which all samples were added was reacted for 2 minutes at 37 degreeC. DOPAchrome produced during the reaction was measured at OD475 nm wavelength.

2. Results

2-1. Analysis of Skin Cell Toxicity by Brassinin

In order to analyze the change in viability caused by brassine, B16F10 cells and HaCaT cells were treated with arbutin (Arbutin, 1 mM), niacinamide (50 uM), and brassine (Brassinin, 10, 20, 40 uM), respectively. . Cells treated with each material were incubated for 3 days and cell viability was analyzed by crystal violet technique.

As a result, as shown in Figure 1, melanogenesis cells (B16F10) and Keratinocyte (HaCaT) cells did not show cytotoxicity. Therefore, it was confirmed that brassinin did not inhibit the cell growth constituting the skin tissue.

2-2. Inhibitory Effect of Brassinin on Tyrosinase

To all groups of tubes was added L-DOPA, a substrate oxidized by tyrosinase enzyme. The group without addition of tyrosinase enzyme isolated from mushrooms in the test tube was set as a control, and the other five groups were added tyrosinase enzyme isolated from mushrooms. Arbutin was added to one of the five test groups, niacinamide was added to the other test group, and bracinin was added to the other two test groups at 20 uM and 40 uM, respectively, and oxidation of L-DOPA by tyrosinase was performed. The color change of the solution generated from was confirmed by absorbance analysis.

As a result, as shown in Figure 2, Arbutin was confirmed to inhibit the activity of about 40% tyrosinase, brassine was confirmed that the inhibitory effect of about 50% tyrosinase activity at 40uM concentration. . Through the above results, it was confirmed that bracinin could inhibit the synthesis of melanin by inhibiting L-DOPA oxidation by tyrosinase.

2-3. Synthesis and Inhibition of Melanin by Brassinin

Melanin-producing cells were pretreated with niacinamide and brassinin, respectively, and allowed to absorb the drug in the cell incubator for 1 hour. After 1 hour, melanin-producing cells were treated with 0.1 mM MSH in four experimental groups except for the control group (see FIG. 3 white graph). After culturing the melanin-producing cells to which MSH and the test substance were added for 3 days, the amount of intracellular melanin was analyzed from the cell extract, and the extracellular melanin was analyzed using the cell culture medium.

As a result, as shown in Figure 3a, it was confirmed that the synthesis of melanin by alpha-MSL (alpha-melanocyte stimulating hormone), a hormone that promotes melanin synthesis, and melanin when niacinamide is added to the MSH treatment group It was confirmed that there was no change in the synthesis of.

Niacinamide is a substance that inhibits the process of melanin synthesis in melanocytes to be transported to keratinocytes. The lack of melanin synthesis in melanocytes is the same as previous results. 40 uM) was confirmed that the melanin synthesis increased by MSH concentration-dependently reduced by brasin.

As a result of confirming whether the melanin synthesized by MSH stimulation is transported out of the melanocytes, as shown in FIG. 3B, the amount of melanin secreted out of the melanocytes was significantly reduced in the experimental group in which niacinamide was added to the MSH treatment group. This result supports the mechanism of inhibiting melanin transport of niacinamide. In the MSH treatment group, the amount of melanin secreted out of the melanocytes was significantly decreased depending on the concentration of brasinin (20, 40uM). It can be said to have the effect of inhibiting the synthesis and secretion of melanin.

So far I looked at the center of the preferred embodiment for the present invention. Those skilled in the art will appreciate that the present invention can be implemented in a modified form without departing from the essential features of the present invention. Therefore, the disclosed embodiments should be considered in descriptive sense only and not for purposes of limitation. The scope of the present invention is shown not in the foregoing description but in the claims, and all differences within the scope equivalent thereto should be construed as being included in the present invention.

Claims (11)

Cosmetic composition for skin whitening comprising brassine (Brassinin) or an acceptable salt thereof represented by Formula 1 below:
[Formula 1]

.
The method of claim 1,
The bracinin or an acceptable salt thereof has a composition which is characterized by inhibiting the production of melanin and inhibitory activity of tyrosinase.
The method of claim 1,
The composition includes cream, softening lotion, nourishing lotion, pack, essence, hair tonic, shampoo, conditioner, hair conditioner, hair treatment, gel, skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisturizing lotion, In any of the formulations selected from the group consisting of nutrition lotion, massage cream, nutrition cream, moisture cream, hand cream, foundation, nutrition essence, sunscreen, soap, cleansing foam, cleansing lotion, cleansing cream, body lotion and body cleanser A composition, which is prepared.
The method of claim 1,
A composition comprising 0.0001 to 10% by weight of the brassinin or an acceptable salt thereof in a total composition content.
A health food composition for skin whitening comprising brassine (Brassinin) represented by Formula 1 or an acceptable salt thereof:
[Formula 1]

.
The method of claim 5,
A composition comprising 0.0001 to 10% by weight of the brassinin or an acceptable salt thereof in a total composition content.
The method of claim 5,
The health food is a health food composition, characterized in that selected from a variety of drinks, meat, sausages, bread, candy, snacks, noodles, ice cream, dairy products, soups, ionic drinks, beverages, alcoholic beverages, gum, tea and vitamin complexes.
A health functional food composition for skin whitening comprising brassine (Brassinin) represented by Formula 1 or an acceptable salt thereof:
[Formula 1]

.
The method of claim 8,
A composition comprising 0.0001 to 10% by weight of the brassinin or an acceptable salt thereof in a total composition content.
The method of claim 8,
The health functional food is a health functional food composition, characterized in that the food in the form of tablets, capsules, pills or liquids.
A composition for skin whitening comprising brassine (Brassinin) represented by Formula 1 or an acceptable salt thereof:
[Formula 1]

.

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